Comparison of dietary essential amino acid requirements determined from group‐housed versus individually‐housed juvenile bluegill,Lepomis macrochirus

Two 60‐day experiments were conducted sequentially to determine (i) lysine requirement of juvenile bluegill, Lepomis macrochirus based on the dose–response method, (ii) requirements for other essential amino acids (EAAs) using whole‐body amino acid profile and (iii) whether differences in growth rates of group‐housed versus individually‐housed bluegills lead to different lysine requirement levels because of the presence and absence, respectively, of social hierarchies. Seven, semi‐purified, experimental diets (isonitrogenous, isocaloric) were prepared to contain graded levels of digestible lysine (10–31 g kg⁻¹). Experiment‐1 involved group‐housed bluegills (approximately 27 g, n = 10 fish/chamber, 4 chambers/diet) whereas experiment‐2 involved individually‐housed bluegills (approximately 30 g, n = 1 fish/chamber, 14 chambers/diet). Fish were fed twice daily to apparent satiation. Bluegill growth responses in both experiments generally improved (P < 0.05, anova ) with increasing dietary lysine levels from 10 to 16 g kg⁻¹, and then levelled off with further increase in lysine level (P > 0.05). Optimal dietary lysine level (digestible basis) was estimated to be 15 g kg⁻¹ based on broken‐line regression analyses of relative growth rate and feed conversion ratio with no differences being observed between the two rearing methods. Determined dietary requirement levels for other EAAs ranged from 2.4 g kg⁻¹ (tryptophan) to 15.3 g kg⁻¹ (leucine).     

大肠杆菌表达系统和酵母表达系统的研究进展

由于DNA重组技术和蛋白组技术的发展与成熟,外源基因表达技术备受关注。其在生物、食品、工业以及医学领域发挥着举足轻重的作用。以原核生物细胞及真核生物细胞作为表达系统,进行目的基因序列的表达以生产蛋白疫苗、核酸疫苗和生物酶制剂等是近年来发酵工业的新型领域。原核和真核表达系统是近年来研究和应用最广泛和最成熟的外源基因表达系统。对近年来关于大肠杆菌表达系统和酵母表达系统的研究进展进行了综述。     

丁酸梭菌对哺乳母猪繁殖性能、免疫和抗氧化功能的影响

本试验旨在研究丁酸梭菌对哺乳母猪免疫和抗氧化的影响,选取健康经产长白母猪32头(胎次和预产期相近),随机分为4组,每组8个重复,每个重复1头。对照组饲喂基础日粮,试验组分别在基础日粮基础上添加250、500、1000 mg/kg丁酸梭菌。预试期7 d,试验期为28 d(产前7 d至21 d断奶)。结果表明:哺乳第1天,1000 mg/kg丁酸梭菌组血清中IgA、IgM和IgG含量均显著高于对照组;哺乳第21天,250、500、1000 mg/kg丁酸梭菌组IgA含量均显著高于对照组,500 mg/kg丁酸梭菌组IgM和IgG含量均为最高;哺乳第1、10、21天,1000 mg/kg丁酸梭菌组IL-6和IL-1β含量较对照组均显著降低;哺乳第1、10、21天,各丁酸梭菌组超氧化物歧化酶活性均显著高于对照组,丙二醛含量均显著低于对照组;哺乳第1、21天,500、1000 mg/kg丁酸梭菌组的过氧化氢酶含量较对照组均显著升高;哺乳第21天,500 mg/kg丁酸梭菌组总抗氧化能力和谷胱甘肽过氧化物酶活性显著升高;哺乳第1天和第21天,1000 mg/kg丁酸梭菌组粪便中大肠杆菌的数量显著低于对照组。由此得出,在妊娠后期和哺乳期饲粮中添加丁酸梭菌可以显著提高母猪免疫功能和抗氧化能力,降低粪便中大肠杆菌的数量,且添加量为500~1000 mg/kg时效果明显。     

广东省养禽场肠球菌耐药性及毒力因子流行分布特征

旨在调查和分析广东省养禽场肠球菌的亚型屎肠球菌和粪肠球菌耐药性及其毒力因子流行分布特征,为控制禽源肠球菌耐药性传播、保障公共卫生安全提供理论依据。作者于2018年从广东省4个养禽场采集肠道样品493份,进行屎肠球菌和粪肠球菌的分离鉴定;采用琼脂二倍稀释法测定肠球菌的最小抑菌浓度（MIC）;PCR方法检测肠球菌的耐药基因和毒力基因。结果显示：1）共分离到125株肠球菌,其中粪肠球菌84株（鸡源66株,鸭源18株）;屎肠球菌41株,均来自鸡肠道样本。2）菌株对四环素、多西环素、红霉素几乎全部耐药,对氟苯尼考和氯霉素的耐药率高达89.60%和74.40%。屎肠球菌耐药率普遍高于粪肠球菌,而粪肠球菌对环丙沙星和利奈唑胺的耐药率高于屎肠球菌;鸭源粪肠球菌对利奈唑胺的耐药率（94%）显著高于鸡源粪肠球菌（39.4%）,屎肠球菌对利奈唑胺均敏感。从鸡分离的1株粪肠球菌对万古霉素耐药。3）耐药基因在屎肠球菌中的检出率高于粪肠球菌,鸭源分离株检出率高于鸡源。耐药基因tetL、fexA、ermB最为流行,检出率均高于90%。其次是optrA基因,检出率为73.60%,poxtA和fexB的检出率均低于20%。在3株鸭源粪肠球菌中检测出cfr基因。4）已检测的毒力基因中efaA的携带率最高,为63.04%（58/92）,其他依次为gelE（54.35%,50/92）、ace（47.83%,44/92）、asa1（44.57%,41/92）。对环丙沙星及高浓度氨基糖苷类耐药的菌株及携带cfr基因的菌株,大多携带agg、asal、gelE或ace。本研究显示养殖场禽源肠球菌耐药严重,鸭源肠球菌对利奈唑胺耐药率高,耐药基因和毒力基因流行且多样,且检测出人医临床重要抗生素耐药基因,应加强对养禽场肠球菌耐药性监测。     

维生素E对蛋鸡产蛋后期产蛋性能和蛋品质的影响

文章旨在研究日粮中不同梯度维生素E添加水平对蛋鸡产蛋后期产蛋性能和蛋品质的影响。试验选择60周龄罗曼白壳商品蛋鸡300羽,试验随机分为5组,添加水平分别为0(对照组),30、60、120、240 mg/kg,每组6个重复,每个重复10羽鸡,预试期1周,正试期8周。结果表明:与对照组相比,蛋鸡产蛋后期日粮中添加120 mg/kg的维生素E可显著提高蛋鸡产蛋后期产蛋性能(P<0.05);与对照组相比,蛋鸡产蛋后期日粮中添加120 mg/kg的维生素E可显著提高蛋壳硬度、蛋壳相对重、蛋黄相对重以及蛋黄颜色(P<0.05),蛋黄总胆固醇含量极显著降低(P<0.01)。综上所述,在蛋鸡产蛋后期日粮中添加120 mg/kg的维生素E可显著改善其产蛋性能和蛋品质。     

鹅大肠埃希菌的分离鉴定及系统进化树分析

为对发病鹅感染致病性大肠埃希菌的临床治疗提供指导并防控致病性大肠埃希菌感染,本研究无菌采集腹泻鹅及病死鹅脾脏、肝脏等组织器官进行病原菌分离培养、染色观察、生化鉴定、致病性试验,对菌株部分基因进行测序、构建遗传进化树并进行药敏试验。结果显示,普通琼脂培养基上生长出灰白色、圆形、凸起、边缘整齐菌落,麦康凯琼脂培养基上长出玫红色、光滑圆润菌落;分离菌株经革兰氏染色,镜下可见革兰氏阴性菌,两端钝圆、大小中等,多呈单个存在的杆状菌;致病性试验表明,该分离菌株对小鼠、雏鹅均有较高致死率;遗传进化树结果显示,分离菌株与患者粪便分离株（GenBank登录号：CP024992.1）同源性最高,达99.5%;体外抑菌试验发现菌株耐药情况较严重,对大多数抗菌药均有较强抗性,仅头孢噻呋对该分离株有较强的抑菌作用,合理用药后病情得到有效控制。本研究为有效防治鹅大肠埃希菌病提供了理论依据,对规模化鹅场防控大肠埃希菌等其他细菌性疾病具有重要价值。     

猪戊型肝炎病毒ORF3蛋白影响HepG2细胞核黄素代谢的lncRNA-mRNA调控网络的鉴定

为初步鉴定并挖掘出猪戊型肝炎病毒(Hepatitis E virus,HEV)ORF3蛋白影响HepG2细胞核黄素代谢信号通路的lncRNA-mRNA调控网络,本试验通过构建腺病毒过表达载体,制备高滴度过表达腺病毒,介导猪 HEV-ORF3在HepG2细胞中实现过表达。Western blotting检测ORF3蛋白过表达成功后,运用lncRNA高通量组学测序,筛选出差异表达的lncRNA并进行靶向差异基因预测,对lncRNA靶向差异基因进行GO功能和KEGG通路富集分析,初步鉴定出与核黄素代谢信号通路相关的lncRNA-mRNA调控网络。Western blotting结果显示,在约为12 ku处出现目的条带,说明成功实现腺病毒介导猪HEV-ORF3在HepG2细胞中过表达。lncRNA高通量组学测序结果显示,共发现102个显著差异表达lncRNAs表达量上调,80个显著差异表达lncRNAs表达量下调。GO功能和KEGG通路富集分析显示,初步挖掘出lncRNA(MSTRG.13995.2)和lncRNA(MSTRG.1960.1)可能是与核黄素代谢信号通路相关的显著差异表达lncRNA,分别通过顺式调控其靶向基因APC-5和FLAD1来影响核黄素代谢信号通路。本试验初步鉴定出lncRNA(MSTRG.13995.2)-APC5和lncRNA(MSTRG.1960.1)-FLAD1可能是影响HepG2细胞核黄素代谢信号通路的lncRNA-mRNA调控网络。     

Effects of periodical application of bioactive peptides derived from cottonseed on performance,immunity, total antioxidant activity of serum and intestinal development of broilers

This experiment aimed to examine the effect of periodical application of bioactive peptides derived from cottonseed (BPC) in comparison with using sub-therapeutic doses of lincomycin and the excessive inclusion of vitamin E on performance, immunity, total antioxidant capacity of serum and intestinal morphology of broiler chickens. A total of 240 one-d-old male broiler chicks with similar initial weight (Ross strain) were randomly assigned to 6 groups (8 chicks/pen): non-treated group (basal diet), basal diet supplemented with 2 mg/kg lincomycin, basal diet supplemented with 50 IU vitamin E, basal diet supplemented with 6 g BPC/kg in starter period, basal diet supplemented with 6 g BPC/kg in starter and grower periods and basal diet supplemented with 6 g BPC/kg throughout the whole experiment. The highest final body weight was obtained in the group supplemented with BPC in starter and grower periods. In the finisher phase, broilers fed the diet containing BPC in the starter period and in the whole trial had significantly (P < 0.05) better feed conversion ratios (FCR). Jejunal villus height was significantly elevated in broilers supplemented with antibiotic (P < 0.001), furthermore it tended to be greater in broilers fed BPC in the starter period. The jejunal villus height-to-crypt depth ratio was significantly (P < 0.01) higher in broilers fed the diet containing antibiotic in comparison to other groups. Humoral immune response against Newcastle disease vaccine tended to be elevated in broilers fed the diet containing BPC in the whole trial (P > 0.05). Broilers supplemented with BPC in starter and grower, and in the whole trial had significantly (P < 0.05) higher antibody titers against sheep red blood cells (SRBC). The highest total antioxidant capacity was obtained in broilers supplemented with the excessive level of vitamin E, furthermore it tended to improve in broilers fed the diet containing BPC in the whole trial. In summary, the results of the study indicated that addition of BPC in broiler diets in the whole trial could improve FCR, immune responses and total antioxidant activity of serum, and BPC could be used in broiler diets as an alternative to in-feed antibiotics.     

Effect of Vitamin E on Bovine Granulosa Cells Apoptosis and Proliferation through Cx43

The aim of this study was to determine the effect of vitamin E on Cx43,the mechanism and function of vitamin E on bovine granulosa cells apoptosis and proliferation.In this study,granulosa cells were isolated from bovine ovary and cultivated in vitro by adding different concentration of vitamin E (0,25,50,100,200 and 500 μmol/L) for 24 h.After cultured,apoptotic cells were detected by FCM,mRNA expression levels of BCL2/BAX、P53 and Cx43 genes were determined by Real-time PCR and cell proliferation was detected by CCK8.The results showed that compared to control group,100 μmol/L vitamin E could significantly inhibit the apoptosis of granulosa cells (P<0.05).Real-time PCR detection results showed that vitamin E significantly changed the mRNA expression levels of BCL2/BAX,P53,Cx43 genes (P<0.05).Vitamin E could significantly improve granulosa cells proliferation when granulosa cells were treated for 24 and 36 h (P<0.05).The results provided a theoretical basis on further analysis for studing the influence mechanism of vitamin E on oocytes development and maturity,and improvement of female animal reproduction by influencing granulosa cells proliferation and apoptosis.     

Prokaryotic Expression and Antigenicity Analysis of Porcine Japenese Encephalitis Virus Envelope Protein Domain Ⅲ

In order to analyze the antigenicity of porcine Japanese encephalitis virus (JEV) E protein domain Ⅲ, which was expressed by pET-28a vector with His-tag and purified through Ni-NTA, the BALB/c mice were immunized with the purified protein.We identified the antigenicity of domain Ⅲ of E protein and the anti-mice and anti-porcine JEV E Ⅲ protein specific antibody titers by SDS-PAGE, Western blotting, indirect ELISA and IFA.SDS-PAGE results showed the expressed target protein existed mainly in the form of inclusion body.Western blotting, ELISA test results showed that the protein had good reactivity with anti-serum.The mice immunized with the purified JEV E Ⅲ protein generated 1×10⁵ anti-JEV E Ⅲ protein specific antibody titers by ELISA, and the porcine immunized with the porcine JEV generated 5.1×10⁴ anti-JEV specific antibody titers.The IFA results showed that JEV E Ⅲ protein anti-serum could identify JEV antigen.The above results showed that the recombinant JEV E Ⅲ had good antigenicity.These results provided important basis for development of diagnostic antigen for JEV.